del Pilar Gomez M, Nasi E
Department of Physiology, Boston University School of Medicine, Boston, Massachusetts 02118, and Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA.
J Neurosci. 1998 Jul 15;18(14):5253-63. doi: 10.1523/JNEUROSCI.18-14-05253.1998.
Visual excitation in rhabdomeric photoreceptors is thought to be mediated by activation of a light-regulated phospholipase C (PLC) and the consequent hydrolysis of phosphatidylinositol bisphosphate. Whereas much attention has been devoted to inositol trisphosphate (IP3) production and intracellular Ca2+ release, little is known about the possible role of the DAG branch in the generation of the light response. We have tested the effect of chemically distinct surrogates of DAG on isolated Lima photoreceptors. Application of the phorbol ester PMA (0.5-10 microM) or the alkaloid (-)-indolactam (20-100 microM) from a holding potential of -50 mV elicited an inward current, several hundred picoamperes in amplitude, accompanied by a pronounced increase in membrane conductance. The stereoisomers 4alpha-PMA and (+)-indolactam were both inactive, arguing for the specificity of the effects. Elevation of cytosolic Ca2+ by intracellular dialysis accelerated this current, whereas chelerythrine antagonized it, suggesting the involvement of PKC. The reversal potential of the membrane current induced by PKC activators was approximately +10 mV; replacement of extracellular Na with impermeant N-methyl-D-glucamine decreased its amplitude and shifted the reversal potential in the negative direction. Stimulation by PMA and (-)-indolactam was accompanied by a pronounced depression of light responsiveness; conversely, steady illumination reduced the size of the current elicited by these PKC activators. Taken together, these results support the notion that the DAG branch of the PLC cascade, in addition to its suggested participation in visual adaptation, may play a role in the activation of the photoresponse or a component thereof, probably in synergy with IP3-mediated Ca2+ release.
横纹肌光感受器中的视觉兴奋被认为是由光调节磷脂酶C(PLC)的激活以及随之而来的磷脂酰肌醇二磷酸的水解介导的。尽管人们对肌醇三磷酸(IP3)的产生和细胞内Ca2+释放给予了很多关注,但对于二酰基甘油(DAG)分支在光反应产生中的可能作用却知之甚少。我们已经测试了化学性质不同的DAG替代物对分离的利马光感受器的影响。从-50 mV的保持电位施加佛波酯PMA(0.5 - 10 microM)或生物碱(-)-吲哚内酰胺(20 - 100 microM)会引发内向电流,幅度为几百皮安,同时膜电导显著增加。立体异构体4α-PMA和(+)-吲哚内酰胺均无活性,这表明了这些效应的特异性。通过细胞内透析升高胞质Ca2+会加速这种电流,而白屈菜红碱则会拮抗它,这表明蛋白激酶C(PKC)参与其中。PKC激活剂诱导的膜电流的反转电位约为 +10 mV;用不透性的N-甲基-D-葡糖胺替代细胞外Na会降低其幅度并使反转电位向负方向移动。PMA和(-)-吲哚内酰胺刺激伴随着光反应性的显著降低;相反,持续光照会减小这些PKC激活剂引发的电流大小。综上所述,这些结果支持了这样一种观点,即PLC级联反应的DAG分支除了其在视觉适应中的推测作用外,可能在光反应或其一个组成部分的激活中发挥作用,可能与IP3介导的Ca2+释放协同作用。